Validation of DNA probes for molecular cytogenetics by mapping onto immobilized circular DNA Metadata

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Title

  • Main Title Validation of DNA probes for molecular cytogenetics by mapping onto immobilized circular DNA

Creator

  • Author: Greulich-Bode, Karin M.
    Creator Type: Personal
  • Author: Wang, Mei
    Creator Type: Personal
  • Author: Rhein, Andreas P.
    Creator Type: Personal
  • Author: Weier, Jingly F.
    Creator Type: Personal
  • Author: Weier, Heinz-Ulli G.
    Creator Type: Personal

Contributor

  • Sponsor: Life Sciences Division
    Contributor Type: Organization

Publisher

  • Name: Lawrence Berkeley National Laboratory
    Place of Publication: Berkeley, California
    Additional Info: Ernest Orlando Lawrence Berkeley National Laboratory, Berkeley, CA (United States)

Date

  • Creation: 2008-12-04

Language

  • English

Description

  • Content Description: Fluorescence in situ hybridization (FISH) is a sensitive and rapid procedure to detect gene rearrangements in tumor cells using non-isotopically labeled DNA probes. Large insert recombinant DNA clones such as bacterial artificial chromosome (BAC) or P1/PAC clones have established themselves in recent years as preferred starting material for probe preparations due to their low rates of chimerism and ease of use. However, when developing probes for the quantitative analysis of rearrangements involving genomic intervals of less than 100kb, careful probe selection and characterization are of paramount importance. We describe a sensitive approach to quality control probe clones suspected of carrying deletions or for measuring clone overlap with near kilobase resolution. The method takes advantage of the fact that P1/PAC/BAC's can be isolated as circular DNA molecules, stretched out on glass slides and fine-mapped by multicolor hybridization with smaller probe molecules. Two examples demonstrate the application of this technique: mapping of a gene-specific {approx}6kb plasmid onto an unusually small, {approx}55kb circular P1 molecule and the determination of the extent of overlap between P1 molecules homologous to the human NF-{kappa}B2 locus. The relatively simple method presented here does not require specialized equipment and may thus find widespread applications in DNA probe preparation and characterization, the assembly of physical maps for model organisms or in studies on gene rearrangements.

Subject

  • Keyword: Tumor Cells
  • Keyword: Chromosomes
  • STI Subject Categories: 59
  • Keyword: Fluorescence
  • Keyword: Genes
  • Keyword: Quality Control
  • Keyword: In-Situ Hybridization
  • Keyword: Plasmids
  • Keyword: Resolution
  • Keyword: Glass
  • Keyword: Dna
  • Keyword: Validation
  • Keyword: Hybridization
  • Keyword: Recombinant Dna

Collection

  • Name: Office of Scientific & Technical Information Technical Reports
    Code: OSTI

Institution

  • Name: UNT Libraries Government Documents Department
    Code: UNTGD

Resource Type

  • Report

Format

  • Text

Identifier

  • Report No.: LBNL-3116E
  • Grant Number: DE-AC02-05CH11231
  • DOI: 10.2172/982916
  • Office of Scientific & Technical Information Report Number: 982916
  • Archival Resource Key: ark:/67531/metadc1013868